Single-use manifold for automated, aseptic transfer of solutions in bioprocessing applications

ABSTRACT

Presteralized manifolds are provided which are designed for sterile packaging and single-use approaches. Disposable tubing and flexible-wall containers are assembled via aseptic connectors. These manifolds interact with at least one remotely controlled pinch valve which engages only the outside surface of the manifold tubing. Such manifold and pinch valve systems can be used in conjuntion with a peristaltic type of pump, which, together with the remotely operated pinch valve, can be operated by a controller which provides automated and accurate delivery of biotechnology fluid in an aseptic environment while avoiding or reducing cleaning and quality assurance procedures.

FIELD OF THE INVENTION

The invention generally relates to the aseptic transfer of solutions outof one or more biological fluid and/or process fluid storage or supplycontainers. Single-use manifold systems carry out transfers needed inbioprocessing applications. With the invention, automated dispensing isaccomplished, preferably in association with one or more remotelycontrolled pinch valves.

BACKGROUND OF THE INVENTION

Good manufacturing practices and governmental regulations are at thecore of any pharmaceutical, biotechnology and biomedical manufacturingprocess or procedure. Such manufacturing processes and procedures aswell as associated equipment must undergo mandated, often lengthy andcostly validation procedures.

For example, the equipment used for the separation and purification ofbiomedical products must, for obvious reasons, meet stringentcleanliness requirements. The cleaning validation of new orre-commissioned purification equipment (such as equipment forpreparative chromatography or tangential flow filtration) may require asmany as 50 test-swabs of exposed surfaces and subsequent biologicalassays of such test-swabs. For a single piece of purification equipment,the associated and reoccurring cost of a single cleaning validation mayreadily exceed multiple thousands of dollars.

To reduce such cleaning validation costs and expenses, and/or to reducethe occasions when cleaning is needed or required, the pharmaceuticaland biotech industries are increasingly employing, pre-sterilized,single-use, plastic tubing and collapsible, plastic bags for solutiontransfer and storage. Sterilization is accomplished by exposing thecomplete tube/bag manifold to gamma irradiation, or to an ethylene oxideatmosphere. The pre-sterilized, aseptically packaged tube/bag manifoldsare commercially available (currently from TC Tech; HyClone; St GobainPerformance Plastics, for example) and are used for the manual transferof solutions. Typically, the solution transfer procedure requires atechnician to operate a peristaltic pump and to manually open and closetube clamps for diverting the solution from the reservoir to the storagebags. Although this procedure reduces the cleaning efforts and cleaningvalidation expense, operator interaction and time still are required,and these approaches are dependent upon operator expertise forconsistent accuracy and precision.

Dispensing approaches having automated features (which can includesensors, monitors and programmable controllers) are generally known.Keys et al. U.S. Pat. No. 5,480,063 and U.S. Pat. No. 5,680,960 describefluid dispensing units which control fluid volumes in conjunction with aclosed loop approach, which these patents suggest can avoid the need forventing. The fluid to be dispensed exits the closed loop apparatusthrough a fill tube, as directed by a controller. Such approaches do notaddress the cleaning needs and/or cleaning validation costs andexpenses, were these types of systems to be used in pharmaceutical andbiotech industries for dispensing, directing, combining or separatingbiological or chemical fluids.

Prior systems can incorporate diaphragm valves, which come into directcontact with the process solution, and these valves are a potentialsource of contamination. Thus diaphragm valves require costly cleaningvalidation procedures.

It has been found that, by proceeding in accordance with the presentinvention, significant cost savings and better performance can berealized in a system which incorporates automated, aseptic manifoldswithin the field of technology which embraces pre-sterilized, single-useplastic tubing and containers having at least one collapsible portion.The components which contact the biological or chemical fluid are eachpresterilized and disposable after use.

SUMMARY OF THE INVENTION

The present invention is directed to manifold units which arepresterilized and disposable, making them single-use units which aresterilized and packaged so as to be usable “off the shelf” and whichthus directly address the problem of tedious and time consuming cleaningand testing at the use site. Multiple embodiments are disclosed. Eachincludes tubing lengths and a plurality of single-use storage orcollection bags, each having multiple inlet and/or outlet passages whichare selectively openable and closeable. The tubing lengths interact withone or more pinch valves which are operable remotely. Remote operationis automated by a controller programmed to carry out proceduresaccording to a selected embodiment.

It is a general object of the present invention to provide improvedsingle-use manifolds for automated, aseptic transfer of solutions inbio-processing or chemical processing applications.

Anther object of the present invention is to provide improved apparatusand method which combine pinch valve use with disposable, sterilizedmanifold dispenser units.

Another object of this invention is to provide improved apparatus andmethod which greatly reduce the expenditure of time and resourcesdevoted to cleaning procedures for transfer equipment used inpharmaceutical and biological industries and laboratories wherecontamination of biological and/or chemical fluids cannot be tolerated.

An aspect of the present invention is to reduce the need for validationprocedures for equipment used in separation and purification of fluidssuch as in conjunction with the preparation, separation and dispensingof biomedical products.

Another aspect of this invention is that it handles cleanlinessrequirements for procedures such as fluid dispensing, preparativechromatography and tangential flow filtration while automating operationthereof.

These and other objects, aspects, features, improvements and advantagesof the present invention will be clearly understood through aconsideration of the following detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

In the course of this description, reference will be made to theattached drawings, wherein:

FIG. 1 is a somewhat schematic illustration of a single-use,presterilized system which is especially suitable for solution transferand collection;

FIG. 2 is an illustration of the single-use system of FIG. 1 inoperative association with pinch values, at least one of which isremotely operable;

FIG. 3 is an illustration of the combination of the features of FIG. 1and FIG. 2, shown with means for use to transfer solution through thesystem;

FIG. 4 is a somewhat schematic illustration of a single-use,presterilized system which is especially suitable for use in automatedpreparative chromatography;

FIG. 5 is an illustration of the single-use system of FIG. 4 inoperative association with pinch valves, at least one of which isremotely operable;

FIG. 6 is an illustration of the combination of features of FIG. 4 andFIG. 5, shown with means for use in transferring solution through thesystem;

FIG. 7 is a somewhat schematic illustration of a single-use,presterilized system which is especially suitable for automatedtangential flow filtration procedures;

FIG. 8 is an illustration of the single-use system of FIG. 7 inoperational association with pinch valves, at least one of which isremotely operable; and

FIG. 9 is an illustration of the combination of the features of FIG. 7and FIG. 8, shown with means for use to transfer solution through thesystem.

DESCRIPTION OF THE PARTICULAR EMBODIMENTS

A system particularly designed for use as an automated, aseptic solutiontransfer system is illustrated in FIGS. 1-3. Fluids processed accordingto this invention are variously referred to herein as biotechnologyfluids, pharmaceutical fluids, chemical fluids, and so forth. These areunderstood to be solutions, liquids, gas-including systems, and thelike. In general, these are referred to herein as biotechnology fluid orfluids.

In the pharmaceutical and biotechnology industries, media preparationdepartments typically prepare the solutions used in a solutionproduction protocal which follows good manufacturing practices. Mediapreparation departments are responsible for maintaining solutionrecipes, preparing and storing buffer solutions and other tasksdemanding consistency and accuracy. For example buffer solutions areprepared in large vats, then pumped through a sterilizing filter, suchas one having a porosity of 0.1μ. Typically such solutions need to befilled into presterilized, single use storage bags for later use. Amedia preparation department may also be responsible for providinginoculating solutions to the operators of a bioreactor. At thecompletion of a bioreactor batch, the reactor broth often is filled intosterile storage bags for later processing.

FIG. 1 shows single-use, presterilized components of the invention.Generally, these disposable components are a manifold and transfertubing assembly and a plurality of bags. A plurality of single-usestorage/collection bags 21, 22, 23 are shown. Each has three tubeconnections. The primary inlet tubing consists of an aseptic connector24 and a manual shut-off clamp 25, each of generally known construction.During solution storage, the aseptic connector is covered with an endcap (not shown) to protect the connector 24 from contamination. Themanual shut-off clamp 25 is closed during solution storage. These areshown on a first tube connection 30.

The second tube connection 26 consists of a short piece of tubingconnected to the bag with a closed manual shut-off clamp. This tubingand clamp arrangement is used to relieve any gas and/or pressurebuild-up inside the bag during the filling operation. The third tubeconnection 27 is identical to the second connection and includes a shortpiece of tubing and a clamp. This can be used as an auxiliary inletand/or outlet for recirculation of the bag contents.

During a typical bag-filling operation, the first and/or last collectionbag can serve the purpose of quality control bags. Often these qualitycontrol bags will be smaller in volume, such as one liter. During theinitial system priming cycle, the first such quality assurance (QA) bagis filled with process solution. At the end of the dispensing cycle whenall of the bags containing the product of the operation, usually largerin volume that the QA bag(s), have been filled, the second QA bag isfilled. The solutions contained in the QA bags are subsequently analyzedfor contamination or for other quality assurance needs.

When the bag-filling process is completed, the manual shut-off clamps oneach bag are closed and the aseptic tube connections are disconnected.During storage, the aseptic connector ends are protected with end caps(not shown).

Turning now to the single-use, sterilized manifold and transfer tubingassembly of FIG. 1, one such unit is generally shown at 28. Thisrepresents a generalized manifold for automated solution transfer. Aninlet end portion 29 of transfer tubing 31 of the unit 28 is forcommunication with a container, such as a vat, of solution, typicallysterile solution. Sterilized manifold and transfer tubing assembly 28 isshown with an optional, in-line pressure sensor 32 and a single-usesterilizing filter 33. An end portion having serially connected endportions are downstream of the illustrated filter 33. By a suitablemovement imparting device, solution moves from the vat or reservoirthrough the sensor 32 (if included) and filter 33 and then is seriallydiverted into the single-use, sterilized storage bags.

FIG. 2 shows a plurality of pinch valves 41, 42, 43 and their respectiverelative positions with respect to the storage bags. Some or all of thevalves can be operated remotely and typically will be pneumatically orelectrically activated. A typical set up will have capacity for up totwelve pneumatically actuated pinch valves or more. A like number ofstorage bags can be accommodated. FIG. 2 shows the relative positions ofthe pinch valves in association with the optional pressure sensor andthe single-use, sterilizing filter. FIG. 3 shows the relative positionof the manifold and transfer tubing assembly 28 with the vat 44 and thepump head of a pump unit 45. Preferably, the pump is a high-accuracy,low-shear peristaltic pump which provides gentle and reproducible bagfilling. An example is a Watson Marlow 620 RE peristaltic pump head.

Access to the storage bags is provided via the pinch valves. The pinchvalves are normally closed and, typical pneumatic pinch valves requirepressurized air (for example 80-100 psi) to open. When such a pinchvalve is pressurized, solution is allowed to enter the storage bag whilethe air in the bag escapes through an integral vent filter. The pinchvalve(s) are pneumatic or electrically operated pinch valves (currentlyavailable from ACRO Associates, Inc). They are installed external to thetubing and are operated by a multi valve controller (currently availablefrom SciLog Inc.), or another computer-based process logic control (PLC)device. The external pinch valves divert the solution inside themanifold without compromising the sterile environment inside the tubing.Diaphragm valves used in other systems are in constant contact with theprocess solution, whereas pinch valves do not contact the processsolution.

The optional, disposable pressure sensor 22 continuously monitors thefilter back pressure. This sensor can provide information to a suitablecontroller to avoid undesired events. For example, a controller canissue an alarm when a safe, user-defined, pressure limit has beenexceeded, indicating that the capacity of the sterilizing filter hasbeen exhausted. Details in this regard are found in U.S. Pat. Nos.5,947,689 and 6,350,382 and in U.S. patent application Publication Ser.No. 2002/0043487, each being incorporated by reference hereinto.

The controller can be a stand-alone unit or be associated with anotherdevice. In a preferred arrangement, the controller is associated withthe pump unit 45. This is shown at 46 in FIG. 3. Whatever form it takes,the controller controls operation of the remotely operable pinchvalve(s). The batch filling rate as well as the batch volume deliveredinto each storage bag is user-programmable via software residing in thecontroller. The controller provides automated bag filling by volume,weight or based on filling time and pump rate.

Typically, a user-determined program will be provided for the automatedfilling of storage bags according to FIGS. 1-3. This is described interms of a SciPro controller of Scilog, Inc., generally described inU.S. Pat. Nos. 5,947,689 and 6,350,382 and U.S. patent applicationPublication Ser. No. 2002/0043487. With these approaches, excessivepressure build-up, as well as associated leaks and bag failures areprevented. For example, when so programmed, the controller will stop allpumping action when a user-defined safe pressure limit is exceeded.

An exemplary solution transfer program for controlling the manifold isas follows. In a SciPro edit mode, the user enters and stores amulti-bag metering program. The following is an example of a simpleprogram to fill three, 20-liter storage bags 21, 22, 23.

Filling Program Example

000 START The following program steps are entered in an edit mode 001 CWMotor Runs Clockwise 002 RUN Motor is tuned “ON” 003 V 100000 PinchValve 41 is Energized, other pinch valves are De-energized 004 RATE:5.01/min Pump Rate 5 liters per minute 005 TIME: 00:04:00 Pump Runs 4minutes, Bag 21 is filled with 20 Liters 006 STOP Pump “Off”, 007 V020000 Pinch Valve 42 is Energized, other valves pinch are De-energized008 TIME: 00:00:02 2 Second Time delay 009 RUN Pump “ON” 010 RATE: 5.0l/min Pump Rate 5 liters per minute 011 TIME: 00:04:00 Pump Runs 4Minutes, Bag 22 is filled with 20 Liters 012 STOP Pump “Off” 013 V003000 Pinch Valve 43 is Energized, other pinch valves are De-energized014 TIME: 00:00:02 2 Second Time Delay 015 RUN Pump “ON” 016 RATE: 5.0l/min Pump Rate 5.0 liters per minute 017 TIME: 00:04:00 Pump Runs 4Minutes, Bag 23 is filled with 20 Liters 018 STOP Pump “Off” 019 V000000 All Pinch Valves are De-energized 020 COUNT: 1 The Program Steps000 to 020 are executed once 021 END

Changes in the RATE and TIME program steps will accommodate any storagebag volume. Additional “RUN” program blocks can be inserted to increasethe number of bags (up to 12 in the example) to be filled. However, ananalogous software program can be generated in which storage bags arefilled based upon either VOLUME or WEIGHT program commands. A scale withan appropriate capacity is required for bag filling by weight. Anoptional scale or load cell 47 can be provided to supply data to thecontroller in this regard. It will be appreciated that this embodimentmeters user-defined volumes of fluid, they automatically switches to thenext empty storage bag to be filled.

A second embodiment, which is generally illustrated in FIGS. 4-6,achieves automated preparative chromatography. In preparativechromatography, process solution containing the bio-molecule of interestis pumped through a column of gel-like particles (stationary phase)suspended in a liquid. The bio-molecule of interest specificallyinteracts (via ion-ion interactions, hydrophobic interactions, sizeexclusion, affinity, for example) with the stationary phase therebyretarding the progress of the bio-molecule through the column. Ideally,other dissolved biomaterials will interact only weakly with thestationary phase and thus will exit the column quickly.

The result is a concentration as well as a separation of thebio-molecule from the rest of the process solution matrix. Theintroduction of an elution buffer will change the local chemicalenvironment of the stationary phase, thereby causing the bio-molecule tobe released and thus able to be collected outside the column in arelatively small volume of elution buffer.

In automated preparative chromatography, the column containing thestationary phase first is washed and/or equilibrated with an appropriatebuffer solution. This wash and/or equilibration cycle is followed by aloading cycle during which the process solution is pumped through thecolumn. The bio-molecule of interest adheres to the stationary phase.The loading cycle can take many hours, depending on the process solutionvolume and pump rate with which the solution is pumped through thecolumn. The loading cycle is followed by a second wash cycle to removeany un-adsorbed biomaterial off the column.

An elution buffer then is introduced to remove the bio-molecule from thecolumn. This removal of the bio-molecule is accomplished either with astep gradient or a linear gradient. After peak collection has beencompleted, the chromatography column is regenerated and re-equilibratedusing appropriate buffer solutions as generally known in the art.

Manifold and transfer tubing assembly 48 represents a generalizedmanifold for automating preparative chromatography procedures. Inoperation, and utilizing the controller system, the exemplarypneumatically controlled pinch valve 51 is pressurized and thus opened,thereby providing access to the wash and/or equilibration buffer bag 54.At a user-definable pump rate, the wash buffer is pumped through adisposable, in-line pressure sensor 55, through a bubble trap (notshown), through the chromatography column 56, and through a detector orUV flow cell 57. On exiting the flow cell, the wash/equilibration bufferis collected in a waste container or bag 58 while pinch valve 49 ispressurized and thus open.

During the loading cycle, pinch valves 51 and 49 are opened/pressurized,while the pinch valves 52, 53 and 59 remain closed. The pump unit 45pumps the process solution through the manifold system 48, the column 56and the flow cell 57 and is collected in the waste container or bag 57.In some chromatography applications, the process solution exiting theflow cell needs to be stored separately in a “process receiving bag”(not shown) for possible re-processing. Another pinch valve (not shown)would provide access to such a “process receiving bag”.

The loading cycle is followed by a wash cycle (valves 51 and 49 areopen/pressurized, all other pinch valves are closed) which carries awayany un-absorbed material from the column to waste. By opening pinchvalves 53 and 49, elution buffer in bag 63 is introduced into the columnand is initially pumped to waste. However, when the signal from the UVdetector 57 exceeds a user-defined value, pinch valve 59 is openedthereby providing access to a peak collection bag 61 while valve 49 isclosed. On the backside of the eluted peak, valve 59 is again closed,while at the same time, valve 49 is opened.

After the material of interest has been collected in bag 61, thechromatographic column 56 requires regeneration and re-equilibration.The column regeneration process is readily automated via access toappropriate buffer solutions (not shown), which are generally as knownin the art. Depending on the underlying chromatographic complexity ofthe application, access to five or six buffer solutions may be required,and these can be provided in their own single-use bags as desired.Similarly, if multiple product peaks are to be collected, additionalpeak collection bag(s) as well as additional pinch valve(s) may have tobe incorporated into manifold and transfer tubing assembly 48.

The single-use, presterilized components of the manifold and transfertubing assembly 48, shown as a feed section, and of a second tube andbag assembly 64 for chromatographed fluid are shown in FIG. 4. Each ofthe single storage/collection bags 54, 62, 63 shown in FIG. 4 has threetube connections. The primary inlet tubing 65 consists of an asepticconnector 66 and a manual shut-off clamp 67. During solution storage,the aseptic connector is covered with an end cap to protect theconnector from contamination. The manual shut-off clamp is closed duringsolution storage.

The second tube and bag assembly 64 consists of a short piece of tubing68 connected to the bag with a closed manual shut-off clamp 68. Thesecond tubing/clamp arrangement 71 is used to relieve any gas and/orpressure build-up inside the bag during the filling operation. The thirdtube connection 72 is identical to the second tubing/clamp arrangement71 and is used as an auxiliary inlet/outlet for recirculation of the bagcontents.

The single-use storage/collection bags 58 and 61 are connected to theremaining tube manifold 72 as shown in FIG. 4 and FIG. 5. FIG. 5 showsthe relative positions of the pinch valves 51, 52, 53, 49 and 59 and theposition of the pressure sensor 55. FIG. 6 shows the insertion of themanifold tubing into the peristaltic pump head 45 as well as connectionsto the chromatography column 56 and the detector 57.

In a typical chromatography application, the single-use storage bags 54(for wash buffer), 52 (for process solution) and 63 (for elution buffer)have been previously filled, for example by using the embodiment ofFIGS. 1-3. When the chromatography run is completed, the manual shut-offclamps on each collection bag 58 (for waste), 61 (for peak collection),and for process receiving (when desired, not shown) are closed, and theaseptic tube connections are disconnected. During storage, the asepticconnector ends are protected with end caps.

Referring further to the SciPro controller programmed for controllingthe manifold arrangement for chromatography, a mode thereof allows entryand storage of a sequence of simple commands, i.e. RUN, RATE, TIME,VOLUME, P LIMIT 1 and Valve States such as V=000000 (all pinch valvesare closed) or V=123456 (all pinch valves are open).

This controller mode is organized in subprogram blocks. The terminatingstatement of a program block can be a “VOLUME”, “TIME”, “P LIMIT D1 (orD2)” or “N LIMIT D1 (or D2)” statement. The statement “P LIMIT D1=5%”reads: “Positive Slope Signal of Detector D1 with a Threshold Value of5% Full Scale (FS)”. See the Chromatography Program Example.

Chromatography Program Example

000 START Start of 1^(st) Wash Cycle 001 CW Clockwise Motor Direction002 RUN Starts Motor 003 RATE 0.25 L/M Pump Rate During Wash Cycle 004 V100050 Wash Buffer 51 Diverted to “Waste” 49 005 VOLUME 4 Minutes, Endof 1^(st) Wash Cycle, TV = 1.0 Liters 1.0 L 006 RATE 1.00 L/M LoadingRate, Start of Loading Cycle 007 V 020050 Process Solution (52) Divertedto “Waste” (49) 008 TIME: 00:02:00 2 Minutes, End of Loading Cycle, TV =3.0 L 009 RATE 0.25 L/M Start of 2^(nd) Wash Cycle 010 V 100050 WashBuffer (51) Diverted to “Waste” (49) 011 VOLUME 4 Minutes, End of 2^(nd)Wash Cycle TV = 1.0 Liter 4.0 L 012 V 003050 Elution Buffer (53)Diverted to “Waste” (49) 013 P LIMIT Threshold Value Detected Start ofPeak D1 = 5% Volume Collection 014 V 003400 Elution Buffer (53) Divertedto “Collect” (59) 015 N LIMIT D1 Threshold Value, End of Peak D1 =10%Volume Collection 016 V 003050 Elution Buffer (53) Diverted to “Waste”(49) 017 VOLUME Elution Volume, End of Elution, 1.0 Liter TV = 5.0 L 018RATE 0.50 L/M Start of 3^(rd) Wash Cycle 019 V 00050 Wash Buffer (51)Diverted to “Waste” (49) 020 TIME 00:02:00 2 Minutes, End of 3^(rd) WashCycle, TV = 6.0 L 021 STOP Pump Stops, 022 V 000000 All V-valves Closed023 END End of Program

For example, in line 014, the SciPro switches from “Waste” to “Collect”when the D1 signal has a positive slope and a value greater than 5% FS(line 013). The statement “N LIMIT D1=10%” reads: “Negative Slope Signalof Detector D1 with a Threshold Value of 10% FS”. In line 016, thecontroller switches from “Collect” to “Waste” when the D1 signal has anegative slope (back side of peak) and a value of 10% FS (line 15).

The user can edit and/or modify the values of: RUN, RATE, TIME, VOLUME,P LIMIT 1, N LIMIT D1 and Valve States at any time during achromatography run. User-designed application programs can be uploadedor downloaded from an external computer at any time by utilizing thecomputer's hyper terminal.

It will be appreciated that, with this embodiment, sequential schedulingof events are achieved. These include sequential scheduling of wash,load and elution cycles. The controller can initiate buffer selection,loading and peak volume collection. Typical in-line concentrationdetectors can be Wedgewood UV and/or pH detectors, which have outputs of4-20 MA outputs which can be monitored simultaneously. A typical pump isa Watson Marlow 620 R peristaltic pump head capable of generating 60 psiat a pump rate of 15 liters per minute.

User-defined detection threshold levels are used for valve switching andpeak volume collection. All solution-handling parameters, such as pumprates, column pressure, and valve positions can be monitored anddocumented in real time and can be printed out or electronicallyarchived.

In a third embodiment, automated tangential flow filtration is carriedout using a modified system designed for this use. Previously referencedU.S. Pat. Nos. 5,947,689 and 6,350,382 and U.S. Published patentapplication Ser. No. 2002/0043487 disclose the automation of tangentialflow filtration (TFF) procedures. These are combined with the use ofdisposable, single-use manifolds, which also include disposable pressuresensors and single-use, collapsible storage bags and the use of remotelyoperated pinch valve(s).

A typical TFF application that utilizes a single-use, pre-sterilizedmanifold is shown in FIGS. 7-9. FIG. 7 shows the disposable,pre-sterilized components, including a tubing filtered fluid sectionhaving a permeate collection bag 81 as well as a process solution bag 82within a filtration flow-through section of the tubing. These areaseptically sealed and in a pre-sterilized (for example, irradiated)package. At the beginning of the TFF application, the permeatecollection bag 81 is empty and deflated and has been asepticallyconnected to the TFF manifold. The process solution bag was previouslyfilled, such as by using the system of FIGS. 1-3. The process solutionbag 82 is placed onto an optional scale 83 and connected aseptically tothe rest of the system. In some applications, weight information can beconveyed to the controller in carrying out the control logic.

The pre-sterilized components of this embodiment are shown in FIG. 7.The permeate collection bag 81 has three tube connections. The primaryinlet tubing 84 consists of an aseptic connector 85 and a manualshut-off clamp 86. During solution storage, the aseptic connector iscovered with an end cap to protect the connector from contamination. Themanual shut-off clamp is closed during solution storage.

The second tube connection consists of a short piece of tubing 87connected to the bag with a closed manual shut-off clamp 88. The secondtubing and clamp arrangement is used to relieve any gas and/or pressurebuild-up inside the bag during the filling operation. The third tubeconnection 89 can be identical to the second tubing and clamparrangement and is used as an auxiliary inlet and outlet forrecirculation of bag contents.

Similarly, the process solution bag 82 has three inlet and/or outlettube connections. The first tube connection 91 is used as an outlet topump solution out of the bag. The second tube connection 92 serves as areturn inlet to accommodate the re-circulated retentate. The third tubeconnection 93 again serves to relieve any excessive gas and/or pressurebuild-up inside the bag.

The permeate collection bag and the process solution bag are connectedto the filtration tube manifold, generally designated at 94 in FIG. 7.FIG. 8 shows the relative positions of the pinch valves 95 and 96 andthe position of three pressure sensors 97, 98, 99. FIG. 9 shows theinsertion of the manifold tubing into the head of the peristaltic pumpunit 45.

Prior to starting the pump unit 45, all of the manual shut-off clampsare opened except those clamps that relieve any gas and/or pressurebuild-up inside the bags. Initially the valve 95 is closed and the valve96 is open, while the pump unit 45 starts to recirculate the solutioncontained in the process solution bag 82 through a tangential flowfilter system 101. The air volume contained in the tubing and tangentialflow filter system 101 ends up in the process solution bag 82 where itis vented to the outside through a sterilizing air filter (not shown).Once the optimal pump recirculation rate has stabilized, pinch valve 95is opened and permeate is collected.

The micro filtration or ultra filtration can be carried out either byconstant rate or by constant pressure. Software programs which aresuitable to automate the filtration process through the use of thecontroller 46 are described in U.S. Pat. Nos. 5,947,689 and 6,350,382and U.S. patent application Publication Ser. No. 2002/00434487.

It will be understood that the embodiments of the present inventionwhich have been described are illustrative of some of the applicationsof the principles of the present invention. Numerous modifications maybe made by those skilled in the art without departing from the truespirit and scope of the invention.

I claim:
 1. A manifold system for biotechnology uses, comprising: amanifold unit which is pre-sterilized and disposable so as to be adaptedfor single-time usage, including: (a) at least one length of tubinghaving at least one inlet end portion, at least one outlet end portion,an outside surface, and an inside surface which is sterilized forpassage of a biotechnology fluid therethrough, (b) at least onesingle-use bag having a primary access port, and (c) an asepticconnector means for operatively connecting said length of tubing withsaid primary access port of the single-use bag; and a plurality of pinchvalves, at least one of which is remotely operable in response to asignal remote from said pinch valve, each said pinch valve engages saidoutside surface of the length of tubing at a discrete locationtherealong, each said pinch valve independently selectively allowing orstopping flow of the biotechnology fluid through said inside surface ofthe length of tubing at said discrete location for that pinch valve; andwherein said system is for automated preparative chromatography, whereinsaid tubing is in at least two sections including a chromatography feedsection and a chromatographed fluid section, wherein said chromatographyfeed section has an outlet and a plurality of serially arranged inletpassageways having one of said aseptic connector means for operableconnection with said single-use bag, wherein said chromatographed fluidsection has an inlet and said outlet end portion of the tubing which hasa plurality of serially arranged outlet passageways having one of saidaseptic connector means for operable connection with said single-usebag, wherein said pinch valves control passage of the biotechnologyfluid from said single-use bags to the chromatography feed section, andwherein another said pinch valve controls passage of the biotechnologyfluid from the tubing chromatographed fluid section to the single-usebag of the chromatographed fluid section.
 2. The manifold system inaccordance with claim 1, wherein said primary access port of thesingle-use bag includes a shut-off clamp.
 3. The manifold system inaccordance with claim 1, wherein said single-use bag further includesaccess port means for releasing gas or pressure build-up from said bag.4. The manifold system in accordance with claim 1, wherein saidsingle-use bag further includes an auxiliary access port.
 5. Themanifold system in accordance with claim 1, wherein said single-use bagfurther includes access port means for releasing gas or pressurebuild-up from said bag and further includes an auxiliary access port. 6.The manifold system in accordance with claim 5, further including ashut-off clamp for said access port means and for said auxiliary accessport.
 7. The manifold system in accordance with claim 1, furtherincluding a single-use sterilizing filter positioned along said lengthof tubing such that the biotechnelogy fluid flows therethrough at alocation upstream of said outlet and portion.
 8. The manifold system inaccordance with claim 1, further including a disposable pressure sensorpositioned along said tubing chromatography feed section such that thebiotechnology fluid flows therethrough at a location upstream of saidoutlet end portion.
 9. The manifold system in accordance with claim 1,further including a chromatography column between said outlet of thechromatography feed section of the tubing and said inlet of thechromatographed fluid section of the tubing.
 10. A manifold system forbiotechnology uses, wherein said system is for automated preparativechromatography, comprising: a manifold unit which is pre-sterilized anddisposable so as to be adapted for single-time usage, including: (a) atleast one length of tubing having at least one inlet end portion, atleast one outlet end portion, an outside surface, and an inside surfacewhich is sterilized for passage of a biotechnology fluid therethrough,(b) a plurality of single-use bags, each having an access port, (c)aseptic connector means for operatively connecting said length of tubingwith said single-use bag, (d) said tubing is in at least two sectionsincluding a chromatography feed section and a chromatographed fluidsection, (e) said chromatography feed section has an outlet and aplurality of serially arranged inlet passageways having one of saidaseptic connector means for operable connection with said single-usebag, and (f) said chromatographed fluid section has an inlet, and saidoutlet end portion of the tubing is on the chromatographed fluid sectionand has a plurality of serially arranged outlet passageways having oneof said aseptic connector means for operable connection with saidsingle-use bag; and a plurality of pinch valves, at least one of whichis remotely operable in response to a signal remote from said pinchvalve, each said pinch valve engages said outside surface of the lengthof tubing at a discrete location therealong, each said pinch valveindependently selectively allowing or stopping flow of the biotechnologyfluid through said inside surface of the length of tubing at saiddiscrete location for that pinch valve, wherein said pinch valvescontrol passage of the biotechnology fluid from said single-use bags tothe chromatography feed section, and wherein another said pinch valvecontrols passage of the biotechnology fluid from the tubingchromatographed fluid section to the single-use bag of thechromatographed fluid section.
 11. The manifold system in accordancewith claim 10, further including a disposable pressure sensor positionedalong said tubing such that the biotechnology fluid flows therethroughat a location upstream of said outlet passageways.
 12. The manifoldsystem in accordance with claim 10, further including a chromatographycolumn between said outlet of the chromatography feed section of thetubing and said inlet of the chromatographed fluid section of thetubing.
 13. The manifold system in accordance with claim 10, wherein atleast one of said single-use bags further includes access port means forreleasing gas or pressure build-up from said bag.
 14. The manifoldsystem in accordance with claim 10, wherein at least one of saidsingle-use bags further includes an auxiliary access port.
 15. Themanifold system in accordance with claim 10, wherein at least one ofsaid single-use bags further includes access port means for releasinggas or pressure build-up from said bag and further includes an auxiliaryaccess port.
 16. The manifold system in accordance with claim 15,further including a shut-off clamp for said access port, for said accessport means, and for said auxiliary access port.
 17. The manifold systemin accordance with claim 10, further including a single-use sterilizingfilter positioned along said tubing such that the biotechnology fluidflows therethrough at a location upstream of said outlet passageways.18. A manifold system for biotechnology uses, wherein said system is fortangential flow filtration, comprising: a manifold unit which ispre-sterilized and disposable so as to be adapted for single-time usage,including: (a) at least one length of tubing having at least one inletend portion, at least one outlet end portion, an outside surface, and aninside surface which is sterilized for passage of a biotechnology fluidtherethrough, (b) a plurality of single-use bags, each having an accessport, one said single-use bag is a process solution bag and another saidsingle-use bag is a permeate collection bag, (c) said tubing is in atleast two sections including a filtration flow-through section and afiltered fluid section, said filtration flow-through section includessaid process solution bag, said filtered fluid section includes saidpermeate collection bag, (d) an aseptic connector for operativelyconnecting said length of tubing with said single-use bag, and (e) adisposable filter between said filtration flow-through section and saidfiltered fluid section, whereby fluid from said process solution bag isfiltered through said disposable filter and is collected in saidpermeate collection bag; and a plurality of pinch valves, at least oneof which is remotely operable in response to a signal remote from saidpinch valve, each said pinch valve engages said outside surface of thelength of tubing at a discrete location therealong, each said pinchvalve independently selectively allowing or stopping flow of thebiotechnology fluid through said inside surface of the length of tubingat said discrete location for that pinch valve.
 19. The manifold systemin accordance with claim 18, wherein said inlet end is within saidfiltration flow-through section and in operative communication with saidprocess solution single-use bag, said filtration flow-through sectionfurther includes a recirculation length having one of said pinch valvesbetween an exit port of said disposable filter and said process solutionsingle-use bag, wherein another said pinch valve is positioned betweensaid disposable filter and said permeate collection single-use bag. 20.The manifold system in accordance with claim 19, further including adisposable pressure sensor positioned along said recirculation length oftubing such that the biotechnology fluid flows therethrough at alocation between said disposable filter and said pinch valve along saidrecirculation length.
 21. The manifold system in accordance with claim18, further including a disposable pressure sensor positioned along saidtubing such that the biotechnology fluid flows therethrough at alocation upstream of said disposable filter.
 22. The manifold system inaccordance with claim 18, further including a disposable pressure sensorpositioned along said filtered fluid length of tubing such that thebiotechnology fluid flows therethrough at a location between saiddisposable filter and said permeate collection single-use bag.
 23. Themanifold system in accordance with claim 18, wherein said single-use bagfurther includes access port means for releasing gas or pressurebuild-up from said bag.
 24. The manifold system in accordance with claim18, wherein said single-use bag further includes an auxiliary accessport.
 25. The manifold system in accordance with claim 18, wherein saidsingle-use bag further includes access port means for releasing gas orpressure build-up from said bag and further includes an auxiliary accessport.
 26. The manifold system in accordance with claim 25, furtherincluding a shut-off clamp for said access port, for said access portmeans, and for said auxiliary access port.
 27. The manifold system inaccordance with claim 18, further including a single-use sterilizingfilter positioned along said tubing such that the biotechnology fluidflows therethrough at a location upstream of said outlet passageways.28. A manifold and pump system for biotechnology uses, wherein saidsystem is for automated preparative chromatography, comprising: amanifold unit which is pre-sterilized and disposable so as to be adaptedfor single-time usage, including: (a) at least one length of tubinghaving at least one inlet end portion, at least one outlet end portion,an outside surface, and an inside surface which is sterilized forpassage of a biotechnology fluid therethrough, (b) a plurality ofsingle-use bags, each having an access port, (c) aseptic connector meansfor operatively connecting said length of tubing with said single-usebag access port, (d) said tubing is in at least two sections including achromatography feed section and a chromatographed fluid section, (e)said chromatography feed section has an outlet and a plurality ofserially arranged inlet passageways having one of said aseptic connectormeans for operable connection with said single-use bag access port, and(f) said chromatographed fluid section has an inlet, and thechromatographed fluid section has and said outlet end portion of thetubing which has a plurality of serially arranged outlet passagewayshaving one of said aseptic connector means for operable connection withsaid single-use bag; a plurality of pinch valves, at least one of whichis remotely operable in response to a signal remote from said pinchvalve, each said pinch valve engages said outside surface of the lengthof tubing at a discrete location therealong, each said pinch valveindependently selectively allowing or stopping flow of the biotechnologyfluid through said inside surface of the length of tubing at saiddiscrete location for that pinch valve, wherein said pinch valvescontrol passage of the biotechnology fluid from said single-use bags tothe chromatography feed section, and wherein another said pinch valvecontrols passage of the biotechnology fluid from the tubingchromatographed fluid section to the single-use bag of thechromatographed fluid section; a chromatography column between saidchromatography feed section and said chromatographed fluid section; anda pump unit which engages said outside surface of the length of tubingat a selected location upstream of said chromatography column.
 29. Amanifold and pump system for biotechnology uses, wherein said system isfor tangential flow filtration, comprising: a manifold unit which ispre-sterilized and disposable so as to be adapted for single-time usage,including: (a) at least one length of tubing having at least one inletend portion, at least one outlet end portion, an outside surface, and aninside surface which is sterilized for passage of a biotechnology fluidtherethrough, (b) a plurality of single-use bags, each having an accessport, one said single-use bag is a process solution bag and another saidsingle-use bag is a permeate collection bag, (c) said tubing is in atleast two sections including a filtration flow-through section and afiltered fluid section, said filtration flow-through section includessaid process solution bag, said filtered fluid section includes saidpermeate collection bag, (d) an aseptic connector means for operativelyconnecting said length of tubing with said single-use bag, and (e) adisposable filter between said filtration flow-through section and saidfiltered fluid section, whereby fluid from said process solution bag isfiltered through said disposable filter and is collected in saidpermeate collection bag; a plurality of pinch valves, at least one ofwhich is remotely operable in response to a signal remote from saidpinch valve, each said pinch valve engages said outside surface of thelength of tubing at a discrete location therealong, each said pinchvalve independently selectively allowing or stopping flow of thebiotechnology fluid through said inside surface of the length of tubingat said discrete location for that pinch valve; and a pump unit whichengages said outside surface of the length of tubing at a selectedlocation upstream of said disposable filter.
 30. An automated manifoldand pump system for biotechnology uses, wherein said system is forautomated preparative chromatography, comprising: a manifold unit whichis pre-sterilized and disposable so as to be adapted for single-timeusage, including: (a) at least one length of tubing having at least oneinlet end portion, at least one outlet end portion, an outside surface,and an inside surface which is sterilized for passage of a biotechnologyfluid therethrough, (b) a plurality of single-use bags, each having anaccess port, (c) aseptic connector means for operatively connecting saidlength of tubing with said single-use bag, (d) said tubing is in atleast two sections including a chromatography feed section and achromatographed fluid section, (e) said chromatography feed section hasan outlet and a plurality of serially arranged inlet passageways havingone of said aseptic connector means for operable connection with saidsingle-use bag, and (f) said chromatographed fluid section has an inletand said outlet end portion of the tubing which has a plurality ofserially arranged outlet passageways having one of said asepticconnector means for operable connection with said single-use bag; aplurality of pinch valves, at least one of which is remotely operable,each said pinch valve engages said outside surface of the length oftubing at a discrete location therealong, each said pinch valveindependently selectively allowing or stopping flow of the biotechnologyfluid through said inside surface of the length of tubing at saiddiscrete location for that pinch valve, wherein said pinch valvescontrol passage of the biotechnology fluid from said single-use bags tothe chromatography feed section, and wherein another said pinch valvecontrols passage of the biotechnology fluid from the tubingchromatographed fluid section to the single-use bag of thechromatographed fluid section; a chromatography column between saidchromatography feed section and said chromatographed fluid section; apump unit which engages said outside surface of the length of tubing ata selected location upstream of said chromatography column; and acontroller which controls operation of said pump unit and of said pinchvalve, said controller having control logic which dictates opening andclosing of said remotely operable pinch valve.
 31. The automated systemin accordance with claim 30, wherein said control logic of thecontroller dictates the rate of pumping of said pump unit.
 32. Theautomated system in accordance with claim 30, wherein said control logicof the controller determines the extent of filling of the single-use bagby processing data monitored by the system to achieve filling of thesingle-use bag by volume, by weight, or by pump rate and filling time.33. The automated system in accordance with claim 30, wherein saidcontrol logic has a loading cycle which activates said pump unit andopens a first and a second said remotely operated pinch valve, saidfirst pinch valve is upstream of said chromatography column and controlsegress of process solution from a container thereof, said second pinchvalve is downstream of said chromatography column and controls access toa first said single-use bag; said loading cycle of the control logicprecedes an elution cycle which opens a third said remotely operatedpinch valve which is upstream of said chromatography column and controlsegress of elution solution from a container thereof and into and throughsaid chromatography column; and said control logic has a peak valuecollection cycle which activates a fourth said remotely operated pinchvalve which is downstream of said chromatography column and controlsaccess of said elution solution into a second said single-use bag. 34.The automated system in accordance with claim 33, wherein said controllogic further includes at least one wash cycle during which said fourthremotely operated pinch valve is closed to deny access to said secondsingle-use bag.
 35. The automated system in accordance with claim 30,further including a detector downstream of said chromatography columnwhich monitors flow out of said chromatography column for a peakcollection value; and wherein said control logic receives peakcollection value data from said detector for use in said peak valuecollection cycle.
 36. An automated manifold and pump system forbiotechnology uses, wherein said system is for tangential flowfiltration, comprising: a manifold unit which is pre-sterilized anddisposable so as to be adapted for single-time usage, including: (a) atleast one length of tubing having at least one inlet end portion, atleast one outlet end portion, an outside surface, and an inside surfacewhich is sterilized for passage of a biotechnology fluid therethrough,(b) a plurality of single-use bags, each having an access port, one saidsingle-use bag is a process solution bag and another said single-use bagis a permeate collection bag, (c) said tubing is in at least twosections including a filtration flow-through section and a filteredfluid section, said filtration flow-through section includes saidprocess solution bag, said filtered fluid section includes said permeatecollection bag, (d) an aseptic connector means for operativelyconnecting said length of tubing with said single-use bag, and (e) adisposable filter between said filtration flow-through section and saidfiltered fluid section, whereby fluid from said process solution bag isfiltered through said disposable filter and is collected in saidpermeate collection bag; a plurality of pinch valves, at least one ofwhich is remotely operable in response to a signal remote from saidpinch valve, each said pinch valve engages said outside surface of thelength of tubing at a discrete location therealong; a pump unit whichengages said outside surface of the length of tubing at a selectedlocation upstream of said disposable filter; and a controller whichcontrols operation of said pump unit and of said pinch valve, saidcontroller having control logic which dictates opening and closing ofsaid remotely operable pinch valve.
 37. The automated system inaccordance with claim 36, wherein said control logic of the controllerdictates the rate of pumping of said pump unit.
 38. The automated systemin accordance with claim 36, wherein said control logic of thecontroller determines the extent of filling of the single-use permeatecollection bag by processing data monitored by the system to achievefilling of the single-use bag by volume, by weight, or by pump rate andfilling time.
 39. The automated system in accordance with claim 36,further including at least one detector positioned along a locationdownstream of said disposable filter for monitoring a parameter of thefluid within said tubing and for transmitting data on the parameter tothe controller, wherein said control logic receives said data from saiddetector and monitors the flow of fluid through the filtration flowthrough section of the tubing until an optimal pump recirculationparameter is achieved, at which time said control logic signals thatsaid filtration flow through section of the tubing is to be blocked byclosing one of said pinch valves and signals that said filtered fluidsection of the tubing is to be unblocked by opening another of saidpinch valves, whereby filtered fluid begins to flow into said single-usepermeate collection bag.
 40. The automated system in accordance withclaim 39, wherein said detector is a pressure sensor, wherein said pumprecirculation parameter is fluid pressure, and wherein said controllogic receives data from said pressure sensor to determine when saidoptimum pump recirculation pressure is achieved.
 41. The automatedsystem in accordance with claim 40, wherein said control logic directsthe pump unit to modify its pumping rate in response to changes inpressure at the pressure sensor so as to maintain a substantiallyconstant selected flow rate imparted to the fluid by the pump unit andthereby assist in achieving said optimum pump recirculation pressure.42. The automated system in accordance with claim 39, wherein saiddetector is a fluid flow rate sensor, wherein said pump recirculationparameter is fluid velocity, and wherein said control logic receivesdata from said fluid flow rate sensor to determine when said optimumpump recirculation fluid velocity is achieved.
 43. The automated systemin accordance with claim 42, wherein said control logic directs the pumpunit to modify its pumping rate in response to changes in flow rate atsaid fluid flow rate sensor so as to maintain a substantially constantselected flow rate imparted to the fluid by the pump unit and therebyassist in achieving said optimum pump recirculation pressure.
 44. Amanifold system for biotechnology uses, comprising: a manifold unitwhich is pre-sterilized and disposable so as to be adapted forsingle-time usage, including: (a) at least one length of tubing havingat least one inlet end portion, at least one outlet end portion, anoutside surface, and an inside surface which is sterilized for passageof a biotechnology fluid therethrough, (b) at least one single-use baghaving a primary access port, and (c) an aseptic connector means foroperatively connecting said length of tubing with said primary accessport of the single-use bag; and a plurality of pinch valves, at leastone of which is remotely operable in response to a signal remote fromsaid pinch valve, each said pinch valve engages said outside surface ofthe length of tubing at a discrete location therealong, each said pinchvalve independently selectively allowing or stopping flow of thebiotechnology fluid through said inside surface of the length of tubingat said discrete location for that pinch valve; and wherein said systemis for tangential flow filtration, wherein one said single-use bag is aprocess solution bag and another said single-use bag is a permeatecollection bag, wherein said tubing is in at least two sectionsincluding a filtration flow-through section and a filtered fluidsection, said filtration flow-through section includes said processsolution bag, said filtered fluid section includes said permeatecollection bag, and further including a disposable filter between saidfiltration flow-through section and said filtered fluid section, wherebyfluid from said process solution bag is filtered through said disposablefilter and is collected in said permeate collection bag.
 45. Themanifold system in accordance with claim 44, wherein said primary accessport of the single-use bag includes a shut-off clamp.
 46. The manifoldsystem in accordance with claim 44, wherein said single-use bag furtherincludes access port means for releasing gas or pressure build-up fromsaid bag.
 47. The manifold system in accordance with claim 44, whereinsaid single-use bag further includes an auxiliary access port.
 48. Themanifold system in accordance with claim 44, wherein said single-use bagfurther includes access port means for releasing gas or pressurebuild-up from said bag and further includes an auxiliary access port.49. The manifold system in accordance with claim 48, further including ashut-off clamp for said access port means and for said auxiliary accessport.
 50. The manifold system in accordance with claim 44, furtherincluding a single-use sterilizing filter positioned along said lengthof tubing such that the biotechnology fluid flows therethrough at alocation upstream of said outlet and portion.
 51. The manifold system inaccordance with claim 44, wherein said inlet end is within saidfiltration flow-through section and in operative communication with saidprocess solution single-use bag, said filtration flow-through sectionfurther includes a recirculation length having one of said pinch valvesbetween an exit port of said disposable filter and said process solutionsingle-use bag, wherein another said pinch valve is positioned betweensaid disposable filter and said permeate collection single-use bag. 52.The manifold system in accordance with claim 44, further including adisposable pressure sensor positioned along said filtration flow-throughsection tubing such that the biotechnology fluid flows therethrough at alocation upstream of said disposable filter.
 53. The manifold system inaccordance with claim 44, further including a disposable pressure sensorpositioned along said filtration flow-through section tubing such thatthe biotechnology fluid flows therethrough at a location between saiddisposable filter and said pinch valve.
 54. The manifold system inaccordance with claim 44, further including a disposable pressure sensorpositioned along said filtered fluid length of tubing such that thebiotechnology fluid flows therethrough at a location between saiddisposable filter and said permeate collection single-use bag.